The Absolute Q® Digital PCR Platform comes in a single optical configuration which is pre-calibrated at the factory. The following compatible dyes may be used in designing custom multiplex assays. Please use the chart below to inform dye selection.
The Absolute Q Digital PCR Platform is pre-calibrated at the factory to support 4-color multiplexing using the most common fluorophores. It is critical to select the correct dyes when designing your multiplexed assay to avoid unexpected results. The above table provides an easy reference to help you find the compatible dyes in each of the 4 channels – Blue, Green, Yellow and Dark Red. In the blue channel, you can use FAM, EvaGreen®, and SYBR® Green. In the green channel, you can use HEX™, VIC®, and Yakima Yellow®. In the yellow channel, you can use TAMRA™ and Cy3™. In the dark red channel, you can use TYE™665, Cy5™, and Quasar670.
Multiplexing increases experimental efficiency and power by providing more data for a given sample run. For instance, multiple targets of interest can be quantified simultaneously against a reference target for a copy number variation assay (CNV). Additionally, the ability to interrogate multiple targets in the same sample is advantageous for detection and quantification of specific single nucleotide variants (SNVs) or single nucleotide polymorphisms (SNPs), which have been implicated in a variety of diseases and clinically relevant phenotypes.
Combinati’s Wastewater Surveillance Kit utilizes all four optical channels to quantify the N1 and N2 gene from SARS-CoV-2 along with Pepper Mild Mottle Virus (PMMoV) for a human fecal control and Bovine Coronavirus (BCoV) as an integrated process control target all in a single reaction. Multiplexing can also be used for genotyping, like in the following publication in Scientific Reports: Development and validation of a 4-color multiplexing spinal muscular atrophy (SMA) genotyping assay on a novel integrated digital PCR instrument.
If you are interested in using other dyes for your application, please reach out to our team and we will be happy to learn more about your needs.